Rapid kinetic studies of ethanol metabolism via the alcohol dehydrogenase-aldehyde hydrogenase pathway and the catalase pathway will be studied in suspensions of hepatocytes to identify the rate-limiting steps, effective enzyme concentration, and to optimize maximal rates of ethanol oxidation via these pathways acting individually and in concert. The effects upon the cellular redox state of varying one or more of these factors will be explored by measurements of the redox state of pyridine nucleotide and flavoprotein in mitochondrial and cytosolic spaces and the role of the redox equilibration systems between these two compartments. A novel dual wavelength-wavelength scanning technique will be devised and will be used to search for intermediates of alcohol oxidation in addition to the catalase, H2O2 intermediate, in hepatocyte suspensions, perfused liver and kidney, and kidney in situ. A novel "tissue freeze-trapping" technique will be employed in the study of the metabolic heterogeneity of the liver, especially to the location of cells in the liver lobule that are maximally participating in ethanol oxidation. The role of catalase in ethanol oxidation will be investigated in red blood cells in order to increase the whole-body clearance of ethanol. Data from these approaches together with those on the alcohol and aldehyde dehydrogenase pathway will be used to provide maximal rates of ethanol clearance with a minimum of metabolic derangement.